ABSTRACT
Reversed-phase chromatography (RPC) seemed to be a technique which was only capable of separating minimal amounts of biological sample per injection or per gram of stationary phase, as has been reported for load capacities using small molecules. Since the problem of fast protein separations at high resolution has been solved by the application of reversed-phase high-performance liquid chromatography (HPLC), the question has now become how to separate large amounts of biological material with good recovery. The major advantage of HPLC with volatile eluents is that sequence and amino acid analysis, as well as gel electrophoresis, can be undertaken directly after evaporation of the salt-free eluents. Ion-exchange chromatography (IEC) seemed to be the only alternative for the separation of large amounts of, e.g., proteins. In the meantime, it has been shown that reversed-phase material can bind amounts of protein that are similar to those which IEC can separate.
